Determination and characterisation of the surface charge properties of the bacteriophage M13 to assist bio-nanoengineering

Passaretti, Paolo; Sun, Yiwei; Dafforn, Timothy R.; Oppenheimer, Pola Goldberg

doi:10.1039/d0ra04086j

Cited by 23 publications

(22 citation statements)

References 42 publications

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“…The structural model of the M13 capsid, analogue of pC89 wild-type vector, has been used to derive the newly rearrangement and electrostatic interactions due to the addition or modification of amino acids in the capsid structure [ 6 ]. In this way, Passaretti et al investigated the surface charge and the surface area of M13 to predict the mechanisms behind of interactions in bio-nanocomponents, finding that the presence of ionizable groups in foreign peptides can drastically change the chemical characteristics of the engineered virus compared to wild-type, playing a role in the resistance to several stresses [ 33 ].…”

Section: Introductionmentioning

confidence: 99%

Role of Phage Capsid in the Resistance to UV-C Radiations

Plano

Franco

Rizzo

et al. 2021

IJMS

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The conformational variation of the viral capsid structure plays an essential role both for the environmental resistance and acid nuclear release during cellular infection. The aim of this study was to evaluate how capsid rearrangement in engineered phages of M13 protects viral DNA and peptide bonds from damage induced by UV-C radiation. From in silico 3D modelling analysis, two M13 engineered phage clones, namely P9b and 12III1, were chosen for (i) chemical features of amino acids sequences, (ii) rearrangements in the secondary structure of their pVIII proteins and (iii) in turn the interactions involved in phage capsid. Then, their resistance to UV-C radiation and hydrogen peroxide (H2O2) was compared to M13 wild-type vector (pC89) without peptide insert. Results showed that both the phage clones acquired an advantage against direct radiation damage, due to a reorganization of interactions in the capsid for an increase of H-bond and steric interactions. However, only P9b had an increase in resistance against H2O2. These results could help to understand the molecular mechanisms involved in the stability of new virus variants, also providing quick and necessary information to develop effective protocols in the virus inactivation for human activities, such as safety foods and animal-derived materials.

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Section: Introductionmentioning

confidence: 99%

Role of Phage Capsid in the Resistance to UV-C Radiations

Plano

Franco

Rizzo

et al. 2021

IJMS

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“…However, this could be associated with the effect of the reduced PEG molecules attached to the surface of M13. We do not exclude the possibility that the presence of PEG could have been interfering with the vibrational modes of the numerous amino and carboxyl groups present on the external surface of the viral capsid 37 . Therefore, these chemical groups were able to show higher absorption values upon PEG removal via isoelectric precipitation (M13 IEP ).…”

Section: Resultsmentioning

confidence: 95%

“…The obtained white pellet consisting of M13 was resuspended in DIW and centrifuged again (5 min at 15,000 rpm) to remove any insoluble fraction. Finally, the concentration of M13 was determined with a UV–Vis spectrophotometer (CaryWinUV by Agilent Technologies), measuring the absorbance at 269 nm and using an extinction coefficient of ε = 3.84 ± 0.06 cm 2 mg −1 37 , 39 .…”

Section: Methodsmentioning

confidence: 99%

“…This can be reduced introducing an additional purification step, i.e., the isoelectric precipitation 36 . This method consists of lowering the pH around the isoelectric point (IEP) of the M13 37 followed by producing the flocculation of the viral particles dispersed in solution and eventually, spinning down to obtain a pellet. Subsequently, the latter can be resuspended in PBS, deionised water (DIW) or any other buffer of choice for specific applications.…”

Section: Introductionmentioning

confidence: 99%

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Improvements in the production of purified M13 bacteriophage bio-nanoparticle

Passaretti

Khan

Dafforn

et al. 2020

Sci Rep

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M13 bacteriophage is a well-established versatile nano-building block, which can be employed to produce novel self-assembled functional materials and devices. Sufficient production and scalability of the M13, often require a large quantity of the virus and thus, improved propagation methods characterised by high capacity and degree of purity are essential. Currently, the ‘gold-standard’ is represented by infecting Escherichia coli cultures, followed by precipitation with polyethylene glycol (PEG). However, this is considerably flawed by the accumulation of contaminant PEG inside the freshly produced stocks, potentially hampering the reactivity of the individual M13 filaments. Our study demonstrates the effectiveness of implementing an isoelectric precipitation procedure to reduce the residual PEG along with FT-IR spectroscopy as a rapid, convenient and effective analytic validation method to detect the presence of this contaminant in freshly prepared M13 stocks.

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“…Like viruses, VLPs have been successfully used in developing vaccines and vaccine adjuvants, and their use in gene therapy and immunotherapy has also been explored (Rohovie et al, 2017;Roldão et al, 2017). Some of those that have shown potential for nucleic acid delivery include bacteriophage-based MS2 (Pan et al, 2012a;Pan et al, 2012b), bacteriophage-based M13 (Yata et al, 2014), animal virus-based hepatitis B virus core (Brandenburg et al, 2005), and plant-based cowpea chlorotic mottle virus (Lam and Steinmetz, 2019).…”

Section: Structure Of Viruses and Genome Protectionmentioning

confidence: 99%

Viral Mimicry as a Design Template for Nucleic Acid Nanocarriers

et al. 2021

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Therapeutic nucleic acids hold immense potential in combating undruggable, gene-based diseases owing to their high programmability and relative ease of synthesis. While the delivery of this class of therapeutics has successfully entered the clinical setting, extrahepatic targeting, endosomal escape efficiency, and subcellular localization. On the other hand, viruses serve as natural carriers of nucleic acids and have acquired a plethora of structures and mechanisms that confer remarkable transfection efficiency. Thus, understanding the structure and mechanism of viruses can guide the design of synthetic nucleic acid vectors. This review revisits relevant structural and mechanistic features of viruses as design considerations for efficient nucleic acid delivery systems. This article explores how viral ligand display and a metastable structure are central to the molecular mechanisms of attachment, entry, and viral genome release. For comparison, accounted for are details on the design and intracellular fate of existing nucleic acid carriers and nanostructures that share similar and essential features to viruses. The review, thus, highlights unifying themes of viruses and nucleic acid delivery systems such as genome protection, target specificity, and controlled release. Sophisticated viral mechanisms that are yet to be exploited in oligonucleotide delivery are also identified as they could further the development of next-generation nonviral nucleic acid vectors.

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Determination and characterisation of the surface charge properties of the bacteriophage M13 to assist bio-nanoengineering

Abstract:
Characterisation of the external surface of bacteriophage M13 using PVIII protein structural data and measuring ζ-potential of the entire virus.

Cited by 23 publications

References 42 publications

Role of Phage Capsid in the Resistance to UV-C Radiations

Role of Phage Capsid in the Resistance to UV-C Radiations

Improvements in the production of purified M13 bacteriophage bio-nanoparticle

Viral Mimicry as a Design Template for Nucleic Acid Nanocarriers

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Determination and characterisation of the surface charge properties of the bacteriophage M13 to assist bio-nanoengineering

Abstract: Characterisation of the external surface of bacteriophage M13 using PVIII protein structural data and measuring ζ-potential of the entire virus.

Cited by 23 publications

References 42 publications

Role of Phage Capsid in the Resistance to UV-C Radiations

Role of Phage Capsid in the Resistance to UV-C Radiations

Improvements in the production of purified M13 bacteriophage bio-nanoparticle

Viral Mimicry as a Design Template for Nucleic Acid Nanocarriers

Contact Info

Product

Resources

About

Abstract:
Characterisation of the external surface of bacteriophage M13 using PVIII protein structural data and measuring ζ-potential of the entire virus.